Regulation of endogenous gene expression in cells using zinc finger proteins
AssigneeSangamo BioSciences, Inc.
InventorsGeorge Norbert Cox, III, Casey Christopher Case, Stephen P. Eisenberg, Eric Edward Jarvis, Sharon Kaye Spratt
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Abstract
The present invention provides methods for modulating expression of endogenous cellular genes using recombinant zinc finger proteins.
Claims85
A method of inhibiting transcription of an endogenous cellular gene in an isolated cell, the method comprising the steps of:
- administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes a first zinc finger protein, wherein said first zinc finger protein is engineered, wherein said polynucleotide sequence is operably linked to a promoter, and wherein the nucleic acid molecule expresses the first zinc finger protein in the cell;
- and
- contacting a first target site in the endogenous cellular gene with the first zinc finger protein, wherein the Kd of the zinc finger protein is less than about 25 nM;
- thereby inhibiting transcription of the endogenous cellular gene by at least about 20%.
A method of inhibiting transcription of an endogenous cellular gene in an isolated cell, the method comprising the steps of:
- administering to the cell a nucleic acid molecule comprising a fusion zinc finger protein encoding nucleic acid operably linked to a promoter, wherein the nucleic acid molecule expresses a fusion zinc finger protein in the cell, and wherein the fusion zinc finger protein comprises six fingers and a regulatory domain;
- and
- contacting a target site in the endogenous cellular gene with the fusion zinc finger protein, wherein the Kd of the fusion zinc finger protein is less than about 25 nM;
- thereby inhibiting transcription of the endogenous cellular gene by at least about 20%.
A method of activating transcription of an endogenous cellular gene in an isolated cell, the method comprising the steps of:
- administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes a first zinc finger protein, wherein said first zinc finger protein is engineered, wherein said polynucleotide sequence is operably linked to a promoter, and wherein the nucleic acid molecule expresses the zinc finger protein in the cell;
- and
- contacting a first target site in the endogenous cellular gene with the first zinc finger protein, wherein the Kd of the zinc finger protein is less than about 25 nM;
- thereby activating transcription of the endogenous cellular gene to at least about 150%.
A method of activating transcription of an endogenous cellular gene in an isolated cell, the method comprising the steps of:
- administering to the cell a nucleic acid molecule comprising a fusion zinc finger protein-encoding nucleic acid operably linked to a promoter, wherein the nucleic acid molecule expresses a fusion zinc finger protein in the cell, and wherein the fusion zinc finger protein comprises six fingers and a regulatory domain;
- and
- contacting a target site in the endogenous cellular gene with the fusion zinc finger protein, wherein the Kd of the fusion zinc finger protein is less than about 25 nM;
- thereby activating transcription of the endogenous cellular gene to at least about 150%.
A method of modulating transcription of an endogenous cellular gene in an isolated cell, the method comprising the steps of:
- administering to the cell a nucleic acid molecule comprising a polynucleotide sequence which encodes a first zinc finger protein, wherein said first zinc finger protein is engineered, wherein said polynucleotide sequence is operably linked to a promoter, and wherein the nucleic acid molecule expresses the zinc finger protein in the cell;
- and
- contacting a first target site in the endogenous cellular gene with the zinc finger protein, thereby modulating transcription of the endogenous cellular gene.
A method of modulating transcription of an endogenous cellular gene in an isolated cell, the method comprising the steps of:
- administering to the cell a nucleic acid molecule comprising a fusion zinc finger protein-encoding nucleic acid operably linked to a promoter, wherein the nucleic acid molecule expresses a fusion zinc finger protein in the cell, and wherein the fusion zinc finger protein comprises six fingers and a regulatory domain;
- and
- contacting a target site in the endogenous cellular gene with the fusion zinc finger protein;
- thereby modulating transcription of the endogenous cellular gene.
Dependencies are inferred from the claim text.